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veterinary

sciences
Communication
Detection of Antibodies to Seven Priority Pathogens
in Backyard Poultry in Trinidad, West Indies
Arianne Brown Jordan 1 ID , Pompei Bolfa 2 , Silvia Marchi 2 , Shakera Hemmings 3 ,
Tashard Major 3 , Rod Suepaul 1 , Lemar Blake 1 and Christopher Oura 1, *
1 Department of Basic Veterinary Sciences, School of Veterinary Medicine, The University of the West
Indies (St. Augustine), Eric Williams Medical Sciences Complex, Mount Hope, Trinidad and Tobago;
arianne.brown@my.uwi.edu (A.B.J.); Rod.Suepaul@sta.uwi.edu (R.S.); Lemar.Blake@sta.uwi.edu (L.B.)
2 Department of Biomedical Sciences, Ross University School of Veterinary Medicine, Basseterre,
Saint Kitts and Nevis; pbolfa@rossvet.edu.kn (P.B.); smarchi@rossvet.edu.kn (S.M.)
3 School of Veterinary Medicine, The University of the West Indies (St. Augustine), Eric Williams Medical
Sciences Complex, Mount Hope, Trinidad and Tobago; shakera.hemmings@my.uwi.edu (S.H.);
tashard.major@my.uwi.edu (T.M.)
* Correspondence: chris.oura@sta.uwi.edu; Tel.: +1-868-645-3232

Received: 15 December 2017; Accepted: 17 January 2018; Published: 20 January 2018

Abstract: Backyard poultry farms in Trinidad and Tobago (T&T) play a vital role in providing
food and income for rural communities. There is currently no information on the presence and
circulation of pathogens in backyard poultry farms in T&T, and little is known in relation to the
potential risks of spread of these pathogens to the commercial poultry sector. In order to address
this, serum samples were collected from 41 chickens on five backyard farms taken from selected
locations in Trinidad. Samples were tested for antibodies to seven priority pathogens of poultry by
enzyme-linked immunosorbent assay (ELISA). Antibodies were detected in 65% (CI 95%: 50–78%)
of the sampled birds for Infectious bronchitis virus (IBV), 67.5% (CI 95%: 52–80%) for Infectious
bursal disease virus (IBDV), 10% (CI 95%: 4–23%) for Newcastle disease virus (NDV), 0% (CI 95%:
0–0%) for Avian influenza virus (AIV), 0% (CI 95%: 0–0%) for West Nile virus (WNV), 31.7% (CI 95%:
20–47%) for Mycoplasm gallisepticum/synoviae and 0% (CI 95%: 0–0%) for Salmonella enterica serotype
Enteritidis. These results reveal the presence and circulation of important pathogens of poultry in
selected backyard farms in Trinidad. The results provide important information which should be
taken into consideration when assessing the risks of pathogen transmission between commercial and
backyard poultry farms, as well as between poultry and wild birds.

Keywords: chickens; Trinidad and Tobago; Avian influenza; infectious bronchitis virus; infectious
bursal disease virus; Newcastle disease virus; West Nile virus; Salmonella enteritidis; Mycoplasma
gallisepticum; Mycoplasma synoviae

1. Introduction
Trinidad and Tobago (T&T) has a large and thriving commercial poultry industry and is
self-sufficient in meat and egg production [1]. It is estimated that over 37 million chickens were
slaughtered for consumption in T&T in 2016 [2]. The country also has a significant number of small
backyard poultry farm operations on both islands, which historically has been a source of food as well
as a hobby for owners [3]. Backyard farms are considered to be small holdings of poultry on family
dwellings with approximately 3–100 birds. The products from backyard farms are commonly used for
home consumption and for small scale selling within local communities. Backyard farmers often rear
mixed poultry flocks along with other animal species. The birds are usually free range and are typically

Vet. Sci. 2018, 5, 11; doi:10.3390/vetsci5010011 www.mdpi.com/journal/vetsci


Vet. Sci. 2018, 5, 11 2 of 6

not vaccinated. In T&T, backyard poultry farms occasionally experience devastating morbidity and
mortality events, which often go unreported and undiagnosed.
Various pathogens have been suspected to cause disease in backyard poultry farms in T&T, but
definitive diagnoses are seldom made. Poultry kept in backyard settings tend to have close interactions
with wild birds, including waterfowl and migratory species, putting them on the frontline for exposure
to pathogens like Avian influenza virus and Newcastle disease virus that are carried by such birds.
Additionally, the lack of farm biosecurity on backyard farms makes the birds more susceptible to certain
pathogens [4]. In this study, a serological approach was used to determine antibody levels for seven
priority pathogens in backyard poultry, chosen based on their zoonotic, veterinary health and public
health importance. Pathogens selected were Avian influenza virus (AIV), Infectious bronchitis virus
(IBV), Infectious bursal disease virus (IBDV), Newcastle disease virus (NDV), West Nile virus (WNV),
Salmonella enterica serotype enteritidis (SE) and Mycoplasma gallisepticum/synoviae (M. gall/synoviae).
The only previous study on backyard poultry in T&T was a poultry production survey conducted
in Northern Trinidad that was published in 1949/1950. This study revealed that ‘peasant’ and backyard
farms experienced confirmed cases of Fowl pox and an undiagnosed ‘mystery disease’ [3]. Although
some work has been conducted into pathogens affecting poultry in backyard settings in the Caribbean,
this is the first such study to take place in T&T for a wide range of pathogens. The objective of this
study was to determine if the selected high priority pathogens of poultry were present and circulating
in backyard poultry in Trinidad and to assess the risks that pathogen circulation in backyard poultry
poses to the commercial poultry sector on the island.

2. Materials and Methods


Chicken samples were collected from January to April 2015 based on convenience and accessibility
of backyard poultry farms in selected locations on the island of Trinidad. A total of 41 samples were
collected from mature (>12 months) unvaccinated birds on five backyard farms. The sampled birds
exhibited no visible signs of clinical disease at the time of sampling. Blood samples were collected in red
top tubes and were transported on ice to the laboratory where the serum was separated and stored at
−80 ◦ C. Serum samples were tested by enzyme-linked immunosorbent assay (ELISA) for the presence
of antibodies against the various pathogens using commercial kits and following manufacturer’s
instructions. Test kits for AIV, IBV, IBDV, NDV and WNV were ID Screen brand kits (IDVet) while both
SE and M. gall/synoviae bacterial kits were Idexx brand kits (Table 1). Sensitivity and specificity levels
of 98% and a 95% confidence level were used to determine confidence intervals (Table 2).

Table 1. ELISA kits used to detect antibodies against various backyard poultry pathogens.

Virus Test kit used Manufacturer


Avian Influenza virus ID Screen®Influenza A Antibody Competition Multi-species ID.Vet
Infectious bronchitis virus ID Screen®Infectious Bronchitis Indirect ID.Vet
Infectious bursal disease virus ID Screen®IBD Indirect ID.Vet
Newcastle disease virus ID Screen®Newcastle Disease Indirect ID.Vet
West Nile virus ID Screen®West Nile Competition Multi-species ID.Vet
Salmonella enteritidis IDEXX SE Ab Test Idexx
Mycoplasma gallicepticum/ synoviae IDEXX MG/MS Ab Test Idexx
Vet. Sci. 2018, 5, 11 3 of 6

Table 2. Antibody detection levels for 7 priority pathogens of backyard chickens in Trinidad,
West Indies.

Positive% (95% CL)


Avian Influenza virus 0% (0,0)
Infectious bronchitis virus 65% (50,78)
Infectious bursal disease virus 67.5% (52,80)
Newcastle disease virus 10% (04,23)
West Nile virus 0% (0,0)
Salmonella enteritidis 0% (0,0)
Mycoplasma gallisepticum/ synoviae 32% (20,47)
CL: Confidence limits.

3. Results
For the viral pathogens, antibodies against IBV, IBDV and NDV were detected at levels of 65%
(CI 95%: 50–78%), 67.5% (CI 95%: 52–80%) and 10% (CI 95%: 04–23%) in the tested bird samples.
No antibodies against AIV and WNV were detected in the birds. For the bacterial pathogens, antibodies
against M. synoviae were detected at levels of 31.7% (CI 95%: 20–47%), whereas no antibodies were
detected for SE. The results from the study are summarized in Table 2. The results indicate the presence
and circulation of IBV, IBDV, NDV and M. gall/synoviae in backyard poultry on the sampled farms in
Trinidad, however antibodies to WNV and SE were not identified to be present.

4. Discussion
This report describes the presence/absence of antibodies to seven priority pathogens of chickens
in a small selection of backyard farms in Trinidad. Due to the small sample size, it was not possible to
interpret the lack of detection of antibodies to Avian Influenza virus, West Nile Virus and Salmonella
enterica. The results do however reveal the presence of antibodies and likely circulation of four
priority pathogens (IBV, IBDV, NDV and M. gall/synoviae) in backyard chickens in Trinidad. Further,
more detailed studies are required in order to fully understand the prevalence of these pathogens in
backyard chickens and the impact of these pathogens on the poultry industry in Trinidad as a whole.
Backyard poultry are thought to be at higher risk than intensively produced poultry of contracting
viruses like AIV, NDV and WNV due to their close contact with wild birds that can act as reservoirs of
these viruses [5–8]. The open layout of backyard farms often results in wild birds interacting closely
with poultry species. This level of interaction is even more significant when there are open water
sources on or near farms, as is often the case. T&T is also along the winter migratory routes for many
migratory bird species, including waterfowl. Many of these migratory birds have been implicated as
carriers of viral pathogens such as AIV, NDV and WNV. One such bird is the blue winged teal, from
which AIV and NDV were isolated in Barbados [9]. In this study no antibodies were detected in the
sampled backyard birds against AIV. A recent study investigating seroprevalence levels for AIV in
domestic layer birds also showed no evidence for circulation of AIV [10]. Continued surveillance of
backyard poultry for the presence of AIV is however important as backyard birds are often in close
contact with potentially infected wild birds, especially during the migration season.
Wild birds are also capable of carrying and transmitting NDV to poultry. The results of this study
indicate that low levels of NDV antibodies (10%) were present in backyard poultry. This relatively low
seroprevalence level in backyard poultry contrasts a recent study carried out in T&T in which up to
80% of unvaccinated layer birds sampled in Trinidad were antibody positive to NDV [10]. These results
suggest that exposure to NDV is lower in backyard poultry compared to commercial layer poultry.
Farm management practices, species and age of birds and exposure to infected birds/environments
could have contributed to the observed difference in seroprevalence levels. As NDV is a highly
infectious virus, more rapid spread would be expected between layer birds kept in close contact
Vet. Sci. 2018, 5, 11 4 of 6

under intensive conditions, than between backyard poultry kept under more extensive conditions.
Given that no clinical signs of disease were noted in the birds sampled in this study, it is likely that
either lentogenic, or possibly attenuated vaccine strains of NDV may be circulating and infecting the
birds. In order to elucidate this, it would be necessary to carry out molecular characterization of the
circulating viruses.
WNV, which is also carried by wild birds and is transmitted through mosquito bites, has also
been detected in the region in both wild birds and domestic poultry. WNV has been detected in many
Caribbean countries including Puerto Rico, Cuba, Dominican Republic, Jamaica and Guadeloupe,
soon after the virus was introduced into the Americas [11–14]. Guadeloupe has carried out several
seroprevalence studies in the period 2002–2004, to look for the presence and spread of the virus.
In these studies, antibodies to WNV were detected in backyard farms as well as on domestic chicken
farms [15,16]. Surveillance activities carried out in Trinidad in 2004 found 2 domestic Muscovy duck
to be seropositive for WNV [17,18], although no virus was isolated. In this current study, no antibodies
to WNV were observed in the sampled backyard birds.
Although less associated with transmission from wild birds, IBV and IBDV are extremely
important viruses affecting poultry worldwide, including in the Caribbean. A recent serological
study revealed high levels of antibodies to both viruses in layer birds in T&T. Antibody detection levels
of close to 100% were reported for both viruses on both islands of Trinidad and Tobago [10]. The lower
prevalence levels for IBV (65%) and IBDV (68%) reported in this study in backyard birds compared to
intensively farmed layer birds may again be a result of multiple factors as described above for NDV.
Both IBV and IBDV are widely vaccinated against in commercial poultry production units in T&T,
however many smaller layer enterprises, as well as backyard farms, do not vaccinate their birds. It is
however unclear whether the IBV and IBDV strains circulating in backyard farms are local field strains
or vaccine derived strains. Molecular characterization of the circulating strain is needed to answer
this question.
M. synoviae and M. gallisepticum can cause symptomatic respiratory tract infections in poultry and
are often associated with IBV and NDV infections [19]. More commonly however, avian mycoplasmosis
presents itself asymptomatically which, though unseen, can cause immunosuppression and poor health
in infected birds [20]. There have been no previous reports of M. synoviae or M. gallisepticum in poultry
in T&T and there are few World Organisation for Animal Health (OIE) reports of its presence and
detection in poultry in the Caribbean [21]. In this study, 31.7% of the backyard birds tested had
antibodies to M. gall/synoviae, indicating that one or both of these bacterial species were present in
birds on the sampled farms. A lower level of detection is expected in backyard compared to intensively
kept poultry, as multiple investigations have reported that smaller less densely populated flocks have
lower levels of infection than larger more densely populated commercial flocks [22]. Further work is
needed to determine if the circulating strains of Mycoplasma in T&T are pathogenic, as a wide range of
pathogenicity and virulence factors are associated with different isolates of M. synoviae [23,24].
The results of this study revealed no antibodies to be present for SE in the backyard poultry that
were sampled. This was surprising given that Salmonella has been detected in poultry houses and pluck
shops throughout Trinidad [25–27], though these detections have been through bacterial enrichment
and culture. Salmonellosis is the primary cause of food borne illness in T&T and the wider Caribbean
and is of great public health concern. SE is one of the most common Salmonella serotypes isolated
from both human and animal sources (poultry, eggs and egg products) in T&T, however there are
multiple other Salmonella serotypes also implicated in causing disease in animals and humans [28,29].
As with Mycoplasma species, farm size is an important factor affecting Salmonella prevalence on farms.
Larger more intensive operations have been found to have higher levels of Salmonella infection and
transmission rates than smaller farms and backyard type farms [26,30].
Vet. Sci. 2018, 5, 11 5 of 6

5. Conclusions
This study confirms the presence of IBV, NDV, IBDV and M. gall/synoviae in unvaccinated backyard
poultry in T&T and provides no evidence for the circulation of WNV and SE in the birds. Due to
the limited sample size, it is not possible to extend the conclusions of this small study to the entire
backyard poultry population in T&T. Nevertheless, the results highlight the presence of four important
pathogens, emphasizing the risks that these pathogens pose to the backyard and commercial poultry
sectors in T&T. The results emphasize the need for changes in farm management and biosecurity
practices in the backyard poultry sector, in order to reduce the exposure of birds to these pathogens.
The selected use of vaccines in this sector may also be recommended. The study also underlines
the need for continuous surveillance and molecular identification and monitoring of disease in the
non-commercial backyard poultry sector, as backyard poultry may act as sentinels for the introduction
of new pathogens into the country and region through wild birds. The backyard poultry sector should
be strongly supported as it plays a highly significant role in contributing to sustainable food security
practices in local communities across the Caribbean region.

Acknowledgments: This work was funded by The University of the West Indies Research, Development and
Impact Fund and The School of Veterinary Medicine Undergraduate Student Project Fund. The authors would
like to thank the poultry farmers in Trinidad who allowed their birds to be sampled and other students who
worked on the project.
Author Contributions: C.O. and R.S. conceived and designed the study; S.H., T.M. and R.S. collected the samples;
A.B.J., L.B., P.B. and S.M. performed the testing; A.B.J. and S.M. analyzed the data; A.J. wrote the paper, which
was revised by CO.
Conflicts of Interest: The authors declare no conflict of interest.

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